Monitoring the distribution of internalized silica nanoparticles inside cells via direct stochastic optical reconstruction microscopy.

Understanding the exact localization of nanoparticles within cell is of particular importance for rational design of high-effective nanomedicines. In the present study, direct stochastic optical reconstruction microscopy (dSTORM) is employed to elucidate the precise localization of nanoparticles within cells owing to its superiority of nanometric resolution, multicolour ability and minimal invasiveness. The localization of the Cy5 labelled mesoporous silica nanoparticles (MSNs-Cy5) in MCF-7 cells are monitored by dSTORM and conventional fluorescence microscopy, respectively. The dSTORM images demonstrate much higher spatial resolution for locating MSNs-Cy5 within cells compared to that of the conventional fluorescence images. Moreover, the distribution of MSNs-Cy5 within three cell lines over time are obtained. For the MCF-7 and HeLa cells, MSNs-Cy5 nanoparticles distribute nearly all around the cytoplasm after 5 h incubation. In contrast, MSNs-Cy5 nanoparticles within NIH 3T3 cells are quite different that they are found to be either attached to or embedded into cell membranes, without penetrating into the cytoplasm. Overall, we provide a practical method to reveal the in situ precise imaging of nanoparticles in cells with nanometric resolution precision. This method may open up new opportunities for organelle-specific targeting drug delivery to achieve maximum therapeutic benefit.

Cell membrane covered polydopamine nanoparticles with two-photon absorption for precise photothermal therapy of cancer.

HYPOTHESIS: In view of the photothermal effect of polydopamine (PDA) nanoparticles and their internal D-π-D structures during assembly, the two-photon excited properties of PDA were studied toward the biomedical application. Further, the PDA molecules were coordinated with Mn2+ and the assembled nanoparticles were covered by cancer cell membranes, the complex system could be used directly for the treatment of cancer with photothermal and chemodynamic therapy. EXPERIMENTS: The two-photon excited PDA-Mn2+ nanoparticles were used for the photothermal therapy combined with chemodynamic therapy. The complexes were coated with cancer cell membranes in order to enhance the tumor homologous efficiency. Multi-modal bioimaging and anti-tumor detections were carried out both in vitro and in vivo. FINDINGS: PDA nanoparticles were demonstrated to have both good two-photon excited fluorescence and photothermal efficiency. The assembled nanoparticles modified with Mn2+ and cancer cell membranes have an obvious targeting and synergetic anti-cancer efficiency. The system creates a simple way for a precise operation with multi-modal imaging function.

Disassembly and reassembly of diphenylalanine crystals through evaporation of solvent.

HYPOTHESIS: Crystalline self-assemblies of diphenylalanine (FF) are since long back considered to be related to Alzheimer's disease. An improved understanding of the mechanism behind the formation of such structures can lead to strategies for investigating the dynamic processes of assembly and disassembly of FF. EXPERIMENT: The assembly, disassembly and reassembly of FF crystals are influenced by the solvent composition and can be triggered by evaporation of solvent. In this work these processes are directly monitored, and the structures obtained are analyzed. FINDINGS: The role of the solvent for assembly, disassembly and reassembly of diphenylalanine crystals has been demonstrated. The initial crystal structure formed via self-assembly of FF monomers can be transformed into needle-like crystals and further to hollow hexagonal microtubes through evaporation of the solvent. It is shown that all the assembly-disassembly processes are spontaneous and driven by thermodynamics. It is also found that some of the crystalline structures exhibit optical waveguiding properties.

Two-photon excited peptide nanodrugs for precise photodynamic therapy.

A novel peptide nanodrug composed of three functional motifs, bis(pyrene), FFVLK and CREKA, was used as a two-photon excited photosensitizer for precise photodynamic therapy (PDT). The system presented excellent two-photon imaging ability, tumor target effect and high reactive oxygen species productivity for improving treatment precision and efficiency in PDT.

Pt@polydopamine nanoparticles as nanozymes for enhanced photodynamic and photothermal therapy.

Polydopamine nanoparticles were used to stabilize a nano-Pt catalyst to relieve tumour hypoxia in photodynamic therapy (PDT). Polydopamine not only provides a platform for carrying nano-Pt and photosensitizers but is also used as a photothermal reagent for photothermal therapy (PTT). The system presented an enhanced anti-tumor therapy effect through a combined PDT and PTT mechanism.

Biological Macrocycle: Supramolecular Hydrophobic Guest Transport System Based on Nanodiscs with Photodynamic Activity.

A biogenic macrocycle-based guest loading system has been developed by the self-assembly of membrane scaffold protein and phospholipids. The resulting 10 nm level transport system can increase the solubility of hydrophobic photodynamic agent hypocrellin B in aqueous medium and exhibited a cellular internalization capacity with substantial photodynamic activity.

Photodynamic Therapy with Liposomes Encapsulating Photosensitizers with Aggregation-Induced Emission.

As a noninvasive treatment, photodynamic therapy (PDT) is a promising strategy against tumors. It is based on photosensitizer (PS)-induced phototoxicity after irradiation. However, most clinically approved PSs will be widely distributed in normal tissues, especially in the skin, where they will induce phototoxicity on exposure to light. Therefore, patients must remain in a dark room for up to several weeks during or after a PDT. Herein, we proposed a strategy of aggregation-induced emission PSs (AIE-PSs) entrapped in liposomes with controlled photosensitization. The AIE-PSs begin to lose their photosensitivity when entrapped in liposomes. After liposomes have carried AIE-PSs into tumor tissues, the AIE-PSs will be released and immediately reaggregate in a targeted area as the liposomes are decomposed. Their photosensitivity can be triggered at turn-on state and induce cytotoxicity. Two different types of AIE molecules were synthesized and entrapped by liposomes, respectively, to verify the PDT features against tumors in vitro and in vivo. The results indicate that, using this strategy, the photosensitivity of AIE-PS can be controlled and PDT can be treated under normal working conditions, not necessarily in a dark room.

A supramolecular hydrophobic guest transport system based on a biological macrocycle.

A protein-based macrocyclic bioactive guest loading system has been developed, which not only provides a stable 10 nm scale lipophilic environment, but also increases the solubility of potent anticancer agent SN38 in its active lactone form in aqueous medium.

Assembled Nanocomplex for Improving Photodynamic Therapy through Intraparticle Fluorescence Resonance Energy Transfer.

In recent years, one of main obstacles in a photodynamic therapy (PDT) process has been that most photosensitizers for PDT are excited by visible light with limited penetrating ability; thus most applications of PDT are for superficial treatments. One of the methods to increase the treatment depth is to introduce a two-photon-active technique into PDT, known as TP-PDT. The difficulty here is to obtain photosensitizers with a large enough two-photon absorption cross-section. In this work, an organic nanocomplex, composed of the two-photon nanoaggregate as the core and photosensitizer as the shell, has been constructed. Photosensitizers could be excited indirectly through a fluorescence resonance energy transfer (FRET) mechanism after the two-photon core was excited by a two-photon laser. The FRET efficiency was extremely high, owing to sufficient energy donors and stable energy acceptors. In this way, a photosensitizer could induce two-photon toxicity for improving the treatment depth in PDT. The nanocomplexes were prepared through a molecular assembly method, which avoided complicated reactions for synthesizing two-photon photosensitizers. The assembly method would expand the selection of photosensitizers and two-photon dyes, and endow traditional photosensitizers with a larger two-photon absorption cross-section for TP-PDT.

Intraparticle FRET for Enhanced Efficiency of Two-Photon Activated Photodynamic Therapy.

Photodynamic therapy (PDT) still faces two main problems on cancer therapy. One is how to improve PDT efficiency against hypoxic environment of tumors. The other one is how to overcome the limit of short wavelength light to increase PDT treatment depth. In this work, an intraparticle fluorescence resonance energy transfer (FRET) platform is designed to address these problems together. The nanoparticles are doped with multicomponents, such as catalase, two-photon dyes, and traditional photosensitizers, with a simple "one-pot" and green method. On the one hand, catalase can catalyze intracellular H2 O2 into O2 and promote PDT efficiency. One the other hand, photosensitizers can be excited indirectly by two-photon lasers through an intraparticle FRET mechanism, which results in deeper tissue penetration for PDT. These properties are verified through the material induced cytotoxicity in light or in dark and in vivo blocking blood-vessel experiment.

An Assembled Nanocomplex for Improving both Therapeutic Efficiency and Treatment Depth in Photodynamic Therapy.

Photodynamic therapy (PDT) shows unique selectivity and irreversible destruction toward treated tissues or cells, but still has several problems in clinical practice. One is limited therapeutic efficiency, which is attributed to hypoxia in tumor sites. Another is the limited treatment depth because traditional photosensitizes are excited by short wavelength light (<700 nm). An assembled nano-complex system composed of oxygen donor, two-photon absorption (TPA) species, and photosensitizer (PS) was synthesized to address both problems. The photosensitizer is excited indirectly by two-photon laser through intraparticle FRET mechanism for improving treatment depth. The oxygen donor, hemoglobin, can supply extra oxygen into tumor location through targeting effect for enhanced PDT efficiency. The mechanism and PDT effect were verified through both in vitro and in vivo experiments. The simple system is promising to promote two-photon PDT for clinical applications.

Nitrogen-doped graphene quantum dots coupled with photosensitizers for one-/two-photon activated photodynamic therapy based on a FRET mechanism.

A novel material with a large two-photon absorption cross-section was conjugated with a typical photosensitizer for inducing a FRET process. The photosensitizer can be excited by a one-/two-photon laser and then induced photo-toxicity in vitro and in vivo. The system presents great potential for improving treatment depth and the precision of traditional photodynamic therapy.

Synthesis, cytotoxicity and antitumour mechanism investigations of polyoxometalate doped silica nanospheres on breast cancer MCF-7 cells.

Polyoxometalates (POMs) have shown the potential anti-bacterial, anti-viral and anti-tumor activities. In order to improve their physiological stability and antitumour activity for medical application, K2Na[AsIIIMo6O21(O2CCH2NH3)3]·6H2O doped silica nanospheres (POM@SiO2) with diameters of ~40 nm have been synthesized by the water-in-oil microemulsion method in this study. The obtained spheres were morphologically uniform nanosized and nearly monodispersed in solution. The nanoparticles had high entrapment efficiency, which was upto 46.2% by the inductively coupled plasma mass spectrometry (ICP-MS) analysis and POMs slowly released from the nanospheres both in the PH 7.4 and 5.5 phosphate buffer saline (PBS) solutions in 60 h. The in vitro MTT assays of particles on MCF-7 cell line (a human breast adenocarcinoma cell line) exhibited enhanced antitumor activity compared to that of plain polyoxometalate. The IC50 value of the POM@SiO2 nanoparticles was 40.0 µg/mL at 24 h calculated by the encapsulated POM concentration, which was much lower comparing to that of 2.0 × 104 µg/mL according to the pure POM. And the SiO2 shells showed low inhibitory effect at the corresponding concentration. Confocal images further indicated the cell morphology changes and necrosis. Flow cytometric analysis showed nanoparticles induced the apoptosis by arresting the cells in S phase and western blot analysis indicated they promoted apoptosis by inhibiting the Bcl-2 protein. Moreover, the study of interactions between human serum albumin (HSA) and the nanoparticles indicated the fluorescence quenching was static, and the nanoparticles were likely to bind to HSA and changed its conformation.

Antileukemic activity of an arsenomolybdate in the human HL-60 and U937 leukemia cells.

The antileukemic activity, mechanisms and serum albumin interactions of an arsenomolybdate, K2Na[AsMo6O21(O2CCH2NH3)3]·6H2O (1), was evaluated in the human leukemia HL-60 and U937 cells. The results indicated that 1 could inhibit the proliferation of both leukemia cell lines in a dose-dependent manner with the 50% lethal concentration (IC50) value of 8.61µM for HL-60 and 14.50µM for U937 at 24h, compare to the positive controls, all-trans retinoic acid (ATRA) with IC50 value of 20.76µM and 14.85µM,and As2O3 with IC50 value of 6.40µM and 8.75µM at 24h, respectively (P<0.05). Furthermore, the anti-leukemia activity of compound 1 might be medicated by arresting the leukemic cells in the G1 phase and inducing apoptosis via caspase-3 and bcl-2 regulatory proteins. Spectroscopic techniques results showed that the fluorescence of human serum albumin was quenched by compound 1, and the quenching mechanism was mainly static quenching. Compound 1 might be a potential medicinal candidate against acute promyelocytic leukemia.

A novel multi-epitope recombined protein for diagnosis of human brucellosis.

BACKGROUND: In epidemic regions of the world, brucellosis is a reemerging zoonosis with minimal mortality but is a serious public hygiene problem. Currently, there are various methods for brucellosis diagnosis, however few of them are available to be used to diagnose, especially for serious cross-reaction with other bacteria. METHOD: To overcome this disadvantage, we explored a novel multi-epitope recombinant protein as human brucellosis diagnostic antigen. We established an indirect enzyme-linked immunosorbent assay (ELISA) based on this recombinant protein. 248 sera obtained from three different groups including patients with brucellosis (146 samples), non-brucellosis patients (82 samples), and healthy individuals (20 samples) were tested by indirect ELISA. To evaluate the assay, a receiver-operating characteristic (ROC) analysis and immunoblotting were carried out using these characterized serum samples. RESULTS: For this test, the area under the ROC curve was 0.9409 (95 % confidence interval, 0.9108 to 0.9709), and a sensitivity of 88.89 % and a specificity of 85.54 % was given with a cutoff value of 0.3865 from this ROC analysis. The Western blot results indicate that it is feasible to differentiate human brucellosis and non-brucellosis with the newly established method based on this recombinant protein. CONCLUSION: Our results obtained high diagnostic accuracy of the ELISA assay which encourage the use of this novel recombinant protein as diagnostic antigen to implement serological diagnosis of brucellosis.

BSA-binding properties and anti-proliferative effects of amino acids functionalized polyoxomolybdates.

Glycine decorated heteropolymolybdates, K2Na[AsMo6O21(O2CCH2NH3)3]·6H2O 1 and K2Na2[γ-Mo8O26(O2CCH2NH3)2]·6H2O 2, have been synthesized and evaluated for in vitro anti-proliferative effects. The identity and high purity of compounds 1 and 2 were confirmed by elemental analysis, FT-IR spectrum, UV-vis spectrum, and X-ray diffraction. Crystal data for 2: triclinic, P-1, a=9.792(2)Å, b=10.077(2)Å, c=10.351(2)Å, α=83.865(4)°, ß=71.110(4)°, γ=62.284(3)°, V=854.3(3)Å(3), Z=1, R(final)=0.0486. The inhibitory effects of 1 and 2 on human non-small cell lung cancer cell line A549 were investigated by MTT assay, nuclear staining, and the flow cytometry. It indicated that compound 1 inhibited the proliferation of A549 cells in a dose-dependent and time-dependent manner, which is more effective than the positive control, 5-fluorouracil (5-FU) (P<0.05). The staining and flow cytometry results showed that compound 1 induced the apoptosis and necrosis of A549 cells and inhibit cell proliferation, which is associated with S-phase arrest. Compound 2 showed a modest activity in a dose-dependent manner. In addition, the interaction between compound 1 and bovine serum albumin (BSA) was evaluated by spectroscopic methods. The results showed that the compound 1 effectively quenched the intrinsic fluorescence of BSA via static quenching and changed the conformation of BSA.